氯喹联合用药体外抗HIV-1作用研究  

The combined anti-HIV-1 effect of chloroquine and antiretroviral drugs in vitro

作　　者：向思颖 陈敏 陈欢[2,3] 罗荣华 杨柳萌[2] 卿晨[1] 郑永唐 

机构地区：[1]昆明医科大学药学院,云南省天然药物药理重点实验室,云南昆明650500 [2]中国科学院昆明动物研究所,云南省活性多肽研究与利用重点实验室/中国科学院动物模型与人类疾病机理重点实验室,云南昆明650223 [3]中国科学院大学,北京100039

出　　处：《药学学报》2018年第2期227-235,共9页Acta Pharmaceutica Sinica

基　　金：国家科技重大专项资助项目(2009ZX09501-029,2014ZX10005-002);国家自然科学基金资助项目(81102483);国家重点研发项目课题(2016YFC1201000);云南省应用基础研究计划(2015FB182).

摘　　要：本文评价氯喹与已上市抗HIV药物联合使用的体外抗HIV-1作用和抑制TLR7激动剂引起的浆细胞样树突状细胞(plasmacytoid dendritic cells,p DC)活化及Ⅰ型干扰素分泌上调的作用。采用MTT法检测氯喹对C8166、TZM-bl、PBMC细胞的毒性。采用荧光素酶活性检测方法、ELISA检测p24抗原测定氯喹分别与雷特格韦(rategrivir,RAL)、恩夫韦肽(enfuvirtide,T-20)、茚地那韦(indinavir,IDV)和依法韦仑(efavirenz,EFV)联合用药抗HIV-1_(ⅢB)、HIV-1_(KM018)活性。流式细胞术检测氯喹分别与RAL、IDV联合使用对细胞p DC活化水平的影响。定量PCR检测氯喹分别与RAL、IDV联合使用对细胞内IFN-α、IFN-βm RNA水平上调的影响。结果表明,氯喹对C8166、TZM-bl和PBMC细胞表现出较低的细胞毒性,半数细胞毒浓度(median cytotoxic concentration,CC_(50))分别为85.02±0.28、73.67±5.10和91.84±4.10μmol·L^(-1)。氯喹分别与RAL、T-20、IDV、EFV联合用药呈中度协同、强协同、相加、中度拮抗效果。氯喹分别与RAL、IDV的联合使用相比于氯喹单药使用对p DC活化水平和IFN-α、IFN-βm RNA表达水平的下调作用无统计学差异。本研究发现氯喹与不同类型的已上市抗HIV药物联合使用呈现不同程度的协同、拮抗或相加抗HIV-1作用。氯喹分别与RAL、IDV联合使用不影响氯喹对TLR7激动剂引起的p DC活化和Ⅰ型干扰素分泌上调的抑制作用。本研究希望能够给氯喹应用于抗HIV治疗提供参考,促进氯喹更广泛地应用于临床研究和治疗中。The study is aimed to evaluate the anti-HIV-1 effect of chloroquine in combination with antihuman immunodeficiency virus （HIV） drugs, and inhibition of plasmacytoid dendritic cells （pDC） activation and type I interferon （IFN-I） production by Toll-like receptor 7 （TLR7） agonist stimulation. We investigated the anti-HIV-1ⅢB, HIV-1KM018 activity of chloroquine and chloroquine combined with rategrivir （RAL）, enfuvirtide （T-20）, indinavir （IDV） and efavirenz （EFV） in vitro by luciferase activity assay system and ELISA method for p24 antigen. We measured the effect of chloroquine on the activation of pDC in combination with RAL and IDV, respectively. Quantitative PCR was used to evaluate the activity of chloroquine in combination with RAL and IDV in the upregulation of interferon （IFN）-α and IFN-β. Chloroquine showed less cytotoxicity to C8166, TZM-bl and PBMC cells, and the 50% cytotoxic concentration values were 85.02 ±0.28, 73.67 ±5.10 and 91.84 ±4.10 μmol·L-1, respectively. The anti-HIV-1ⅢB activity of chloroquine combination with RAL, T-20, IDV and EFV were moderate in synergy, strong in synergy, additive and moderate antagonism, respectively. The anti-HIV-1KM018 activity of chloroquine in combination with RAL, IDV were moderate synergy, minor synergy. There was no significant difference between the chloroquine monotherapy and chloroquine combined with RAL, IDV in the down-regulation of pDC activation and IFN-α, IFN-β expression levels. We have found that chloroquine combined with different anti-HIV drugs represent different degrees of synergism, antagonism or additive anti-HIV-1 effect. Chloroquine in combination with RAL and IDV did not have influence on the inhibitory effect of chloroquine on pDC activation and type I interferon secretion induced by TLR7 agonist. The results suggest that chloroquine may be used to enhance the therapeutic activities of anti-HIV medicines.

关 键 词：氯喹 人类免疫缺陷病毒-1 联合用药 浆细胞样树突状细胞活化 

分 类 号：R966[医药卫生—药理学]